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ENDOMETRIUM Publication-derived

Long-term, hormone-responsive organoid cultures of human endometrium in a chemically-defined medium

Source Turco et al., 2017 · University of Cambridge, Department of Pathology · 10.1038/ncb3516

👤 Margherita Y. Turco, Lucy Gardner, Jasmine Hughes, Tereza Cindrova-Davies, Maria J. Gomez, Lydia Farrell, Michael Hollinshead, Steven G.E. Marsh, Jan J. Brosens, Hilary O. Critchley, Benjamin D. Simons, Myriam Hemberger, Bon-Kyoung Koo, Ashley Moffett, Graham J. Burton ⏱ 14 days 📋 6 phases 🧫 Human endometrial epithelial cells (non-pregnant endometrium and decidua)

Abstract

This protocol establishes long-term, genetically stable 3D organoid cultures from human non-pregnant endometrium and decidua in a chemically-defined expansion medium. The organoids recapitulate endometrial glandular features in vivo, respond to sex hormones, and can differentiate into ciliated and secretory cells upon pregnancy hormone stimulation, providing a model system for studying implantation, early pregnancy, and endometrial pathologies.

Cell source
Human endometrial epithelial cells (non-pregnant endometrium and decidua)
Application
Disease modeling and developmental study of endometrial function, implantation, and early pregnancy

Protocol overview

19 steps across 6 phases

Tissue Processing and Gland Isolation Day 0
  1. 1 Tissue chopping and enzymatic digestion
  2. 2 Filtration and gland collection
Organoid Initiation and Plating Day 0–1
  1. 1 Preparation of Matrigel suspension
  2. 2 Plating in Matrigel droplets
  3. 3 Overlay with Expansion Medium (ExM)
Organoid Expansion and Maintenance Day 1–14+ (passaging every 7–10 days)
  1. 1 Medium refreshment
  2. 2 Manual passaging of organoids
  3. 3 Long-term expansion monitoring
Hormonal Differentiation – Ovarian Hormone Treatment Day 4–10
  1. 1 Estrogen priming
  2. 2 Progesterone and cAMP co-stimulation
  3. 3 Collection for analysis
Pregnancy Signal Stimulation Day 4–12
  1. 1 Preparation of Differentiation Medium (DM)
  2. 2 Addition of pregnancy hormones
  3. 3 Extended differentiation culture
  4. 4 Collection for analysis
Single-Cell Cloning and Clonogenic Assays Day 0–14
  1. 1 Preparation of single-cell suspension
  2. 2 Limiting dilution plating
  3. 3 Scoring organoid formation
  4. 4 Expansion and characterization of clonal lines

Full SOP

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Attribution

This SOP was authored by Organthis based on the published method in Turco et al., 2017. The originating laboratory holds no rights in this SOP and has not endorsed it unless marked Verified.

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